Heterologous expression of neisserial proteins

Abstract

Alternative and improved approaches to the heterologous expression of the proteins of Neisseria meningitidis and Neisseria gonorrhoeae are disclosed. These approaches typically affect the level of expression, the ease of purification, the cellular localization, and/or the immunological properties of the expressed protein.

1 . A method of inducing a bactericidal immune response in an animal, comprising administering to the animal a composition comprising a non-lipidated polypeptide, which comprises an amino acid sequence having greater than 99% sequence identity to the amino acid sequence of ΔG741 from  Neisseria meningitidis  strain MC58, wherein the non-lipidated polypeptide is present in an immunologically effective amount that is effective to elicit bactericidal antibodies against a  Neisseria meningitidis  serogroup B strain in the animal. 
     
     
         2 . The method according to  claim 1 , wherein the immunologically effective amount is at least 20 μg. 
     
     
         3 . The method according to  claim 2 , wherein the immunologically effective amount is 50-200 μg. 
     
     
         4 . The method according to  claim 2 , wherein said composition additionally comprises an effective amount of an adjuvant. 
     
     
         5 . The method according to  claim 4 , wherein the adjuvant is aluminum hydroxide or aluminum phosphate. 
     
     
         6 . The method according to  claim 2 , wherein said non-lipidated polypeptide does not comprise an N-terminal amino acid residue site for lipidation. 
     
     
         7 . The method according  claim 2 , further comprising a pharmaceutically acceptable carrier, adjuvant, diluent or buffer. 
     
     
         8 . The method according to  claim 2 , wherein the non-lipidated polypeptide elicits a bactericidal immune response against a heterologous  Neisseria meningitidis  strain in the animal. 
     
     
         9 . The method according to  claim 2 , consisting essentially of the non-lipidated polypeptide. 
     
     
         10 . The method according to  claim 2 , wherein the composition does not comprise a protein having an amino acid sequence of greater than 80% sequence identity to SEQ ID NO: 2534 in WO99/57280. 
     
     
         11 . The method according to  claim 2 , wherein the non-lipidated polypeptide is a recombinant polypeptide. 
     
     
         12 . The method according to  claim 2 , wherein the non-lipidated polypeptide is a fusion polypeptide. 
     
     
         13 . The method according to  claim 2 , wherein the non-lipidated polypeptide is a purified polypeptide. 
     
     
         14 . The method according to  claim 2 , wherein the non-lipidated polypeptide is conjugated to a carrier. 
     
     
         15 . The method according to  claim 2 , wherein the non-lipidated polypeptide was expressed in  E. coli.    
     
     
         16 . The method according to  claim 1 , wherein the composition comprises
 (a) the non-lipidated polypeptide; and   (b) an immunostimulatory effective amount of an aluminum hydroxide adjuvant.   
     
     
         17 . The method according to  claim 16 , wherein the immunologically effective amount of the non-lipidated polypeptide is at least 20 μg. 
     
     
         18 . The method according to  claim 16 , wherein the immunologically effective amount of the non-lipidated polypeptide is 50-200 μg. 
     
     
         19 . The method according to  claim 16 , wherein said non-lipidated polypeptide does not comprise an N-terminal amino acid residue site for lipidation. 
     
     
         20 . The method according to  claim 16 , further comprising a pharmaceutically acceptable carrier, diluent or buffer. 
     
     
         21 . The method according to  claim 16 , wherein the non-lipidated polypeptide elicits a bactericidal immune response against a heterologous  Neisseria meningitidis  strain in the animal. 
     
     
         22 . The method according to  claim 16 , consisting essentially of (a) and (b). 
     
     
         23 . The method according to  claim 16 , wherein the composition does not comprise a protein having an amino acid sequence of greater than 80% sequence identity to SEQ ID NO: 2534 of WO99/57280. 
     
     
         24 . The method according to  claim 16 , wherein the non-lipidated polypeptide is a recombinant polypeptide. 
     
     
         25 . The method according to  claim 16 , wherein the non-lipidated polypeptide is a fusion polypeptide. 
     
     
         26 . The method according to  claim 16 , wherein the non-lipidated polypeptide is a purified polypeptide. 
     
     
         27 . The method according to  claim 16 , wherein the non-lipidated polypeptide is conjugated to a carrier. 
     
     
         28 . The method according to  claim 16 , wherein the non-lipidated polypeptide comprises the amino acid sequence of ΔG741 from  Neisseria meningitidis  strain MC58. 
     
     
         29 . The method according to  claim 16 , wherein the non-lipidated polypeptide was expressed in  E. coli.